miRNAs differentially expressed in HepG2 cells after 5 hours of culture with grape seed proanthocyanidin extract.

<p>The fold change value and the standard deviations obtained by QRT-PCR is presented in parentheses.</p

miRNAs differentially expressed in HepG2 cells after 5 hours of culture with cocoa proanthocyanidin extract.

<p>The fold change value and the standard deviations obtained by QRT-PCR is presented in parentheses.</p

Validated target genes for the miRNAs differentially expressed in response to both grape seed proanthocyanidin extract and epigallocatechin gallate.

<p>(1): from the miRWalk database (<a href="http://www.umm.uni-heidelberg.de/apps/zmf/mirwalk/" target="_blank">http://www.umm.uni-heidelberg.de/apps/zmf/mirwalk/</a>).</p><p>(2): from the KEGG (<a href="http://www.genome.jp/kegg/" target="_blank">http://www.genome.jp/kegg/</a>) and NCBI-Gene (<a href="http://www.ncbi.nlm.nih.gov" target="_blank">http://www.ncbi.nlm.nih.gov</a>) databases.</p

Selection of validated target genes for miR-30b* grouped by pathway.

<p>(1): from the KEGG (<a href="http://www.genome.jp/kegg/" target="_blank">http://www.genome.jp/kegg/</a>) and BioCarta (<a href="http://www.biocarta.com/" target="_blank">http://www.biocarta.com/</a>) pathway databases.</p><p>(2): from the miRWalk database (<a href="http://www.umm.uni-heidelberg.de/apps/zmf/mirwalk/" target="_blank">http://www.umm.uni-heidelberg.de/apps/zmf/mirwalk/</a>).</p><p>(3): from the KEGG (<a href="http://www.genome.jp/kegg/" target="_blank">http://www.genome.jp/kegg/</a>) and NCBI-Gene (<a href="http://www.ncbi.nlm.nih.gov" target="_blank">http://www.ncbi.nlm.nih.gov</a>) databases.</p

miRNAs differentially expressed in HepG2 cells after 5 hours of culture with epigallocatechin gallate.

<p>The fold change value and the standard deviations obtained by QRT-PCR is presented in parentheses.</p

Table_3_The Exposure to Different Photoperiods Strongly Modulates the Glucose and Lipid Metabolisms of Normoweight Fischer 344 Rats.doc

<p>Seasonal variations in day length trigger clear changes in the behavior, growth, food intake, and reproductive status of photoperiod-sensitive animals, such as Fischer 344 rats. However, there is little information about the effects of seasonal fluctuations in day length on glucose and lipid metabolisms and their underlying mechanisms in this model. To gain knowledge on these issues, three groups of male Fischer 344 rats were fed with a standard diet and exposed to different photoperiods for 14 weeks: normal photoperiod (L12, 12 h light/day), long photoperiod (L18, 18 h light/day), and short photoperiod (L6, 6 h light/day). A multivariate analysis carried out with 239 biometric, serum, hepatic and skeletal muscle parameters revealed a clear separation among the three groups. Compared with L12 rats, L6 animals displayed a marked alteration of glucose homeostasis and fatty acid uptake and oxidation, which were evidenced by the following observations: (1) increased circulating levels of glucose and non-esterified fatty acids; (2) a sharp down-regulation of the phosphorylated Akt2 levels, a downstream post-receptor target of insulin, in both the soleus and gastrocnemius muscles; (3) decreased expression in the soleus muscle of the glucose metabolism-related microRNA-194 and lower mRNA levels of the genes involved in glucose metabolism (Irs1, soleus, and Glut2, liver), β-oxidation (Had and Cpt1β, soleus) and fatty acid transport (Cd36, soleus, and liver). L18 animals also displayed higher blood glucose levels than L12 rats and profound changes in other glucose and lipid metabolism-related parameters in the blood, liver, and skeletal muscles. However, the mechanisms that account for the observed effects were less evident than those reported in L6 animals. In conclusion, exposure to different photoperiods strongly modulated glucose and lipid metabolisms in normoweight rats. These findings emphasize the relevance of circannual rhythms in metabolic homeostasis regulation and suggest that Fischer 344 rats are a promising animal model with which to study glucose- and lipid-related pathologies that are influenced by seasonal variations, such as obesity, cardiovascular disease and seasonal affective disorder.</p

Table_2_The Exposure to Different Photoperiods Strongly Modulates the Glucose and Lipid Metabolisms of Normoweight Fischer 344 Rats.doc

<p>Seasonal variations in day length trigger clear changes in the behavior, growth, food intake, and reproductive status of photoperiod-sensitive animals, such as Fischer 344 rats. However, there is little information about the effects of seasonal fluctuations in day length on glucose and lipid metabolisms and their underlying mechanisms in this model. To gain knowledge on these issues, three groups of male Fischer 344 rats were fed with a standard diet and exposed to different photoperiods for 14 weeks: normal photoperiod (L12, 12 h light/day), long photoperiod (L18, 18 h light/day), and short photoperiod (L6, 6 h light/day). A multivariate analysis carried out with 239 biometric, serum, hepatic and skeletal muscle parameters revealed a clear separation among the three groups. Compared with L12 rats, L6 animals displayed a marked alteration of glucose homeostasis and fatty acid uptake and oxidation, which were evidenced by the following observations: (1) increased circulating levels of glucose and non-esterified fatty acids; (2) a sharp down-regulation of the phosphorylated Akt2 levels, a downstream post-receptor target of insulin, in both the soleus and gastrocnemius muscles; (3) decreased expression in the soleus muscle of the glucose metabolism-related microRNA-194 and lower mRNA levels of the genes involved in glucose metabolism (Irs1, soleus, and Glut2, liver), β-oxidation (Had and Cpt1β, soleus) and fatty acid transport (Cd36, soleus, and liver). L18 animals also displayed higher blood glucose levels than L12 rats and profound changes in other glucose and lipid metabolism-related parameters in the blood, liver, and skeletal muscles. However, the mechanisms that account for the observed effects were less evident than those reported in L6 animals. In conclusion, exposure to different photoperiods strongly modulated glucose and lipid metabolisms in normoweight rats. These findings emphasize the relevance of circannual rhythms in metabolic homeostasis regulation and suggest that Fischer 344 rats are a promising animal model with which to study glucose- and lipid-related pathologies that are influenced by seasonal variations, such as obesity, cardiovascular disease and seasonal affective disorder.</p

Multivariate modelling of ¹N-NMR, GC/MS and LC/ESI-TOF MS data.

<p>(<b>A</b>) PC1/PC2 scatter scores plot and (<b>B</b>) PC1 loading bar plot of PCA calculated on the within-subject matrix derived from the MSCA modelling of the 32 selected spectral regions identified in the ¹H-NMR CPMG serum spectra of untreated (red markers) and treated (green markers) PCOS patients. (<b>C</b>) PC1/PC2 scatter scores plot of PCA calculated on the within-subject matrix derived from the MSCA modelling of GC/MS data. Boxplots of (<b>D</b>) azelaic acid and (<b>E</b>) 1,2-proanediol, the two metabolites corresponding to the most discriminating features along the corresponding PC1 loadings bar plot. (<b>F</b>) PC1/PC2 scatter scores plot of PCA calculated on the within-subject matrix derived from the MSCA modelling of LC/ESI-TOF MS data. Boxplots of (<b>G</b>) 9- and 13-HODE, and (<b>H</b>) caprylic acid, the two metabolites corresponding to the most discriminating features along the PC1 loadings bar plot. P-values derived from Wilcoxon rank-summed pair-matched comparison of untreated and treated PCOS patients. Mean ± sem of the percentage of variation are also indicated.</p

CPMG ¹H NMR spectra of a representative PCOS patient's serum.

<p>(<b>A</b>) Comparative spectra at baseline and after 30 months of Pio/Flu/Met polytherapy. CPMG spin echo experiment allows filtering broad signals of lipid and lipoproteins enhancing low-molecular weight metabolites such as amino acids, lactate and intermediate metabolites. The inset displays an expanded δ (0.75–1.5 ppm) spectral region showing two unidentified resonances characteristic of the serum spectra of untreated PCOS patients: a broad singlet arising at 0.85 ppm and three peaks centered at 1.18 ppm. (<b>B</b>) CPMG ¹H-NMR spectra of the same untreated PCOS patient shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029052#pone-0029052-g002" target="_blank">Figure 2A</a> before and after cold acetone precipitation. After acetone precipitation the three signals centered at 1.18 ppm were depleted, confirming the occurrence of oxidized lipoprotein-related structures in the serum of PCOS women.</p

Correlation between IMT and azelaic acid levels in serum.

<p>Positive significant correlation (r = 0.92, p = 3.96×10⁻⁷) between carotid IMT values and azelaic acid levels. Azelaic acid levels were calculated as the ratio of the fragmentation peak of azelaic acid at m/z = 317 (retention time = 16.38 min) and the peak area of the internal standard. Red and green dots represent values of untreated and treated PCOS patients respectively.</p